ES2271996T3 - METHOD TO ADMINISTER A COMPOSITION OF VIABLE MICROORGANISMS TO CORRAL BIRDS. - Google Patents

Abstract

The invention relates to a method for the administration to birds of a composition based on viable microorganisms which consists in administering a composition based on viable microorganisms that includes viable milk bacteria of the genus Lactobacilus reuteri and Lactobacilus johnsonii in a state of a newborn bird ; and the administration of viable microorganisms that includes a viable microorganism belonging to Bacilus subtilis.

Description

Method for administering a composition of viable microorganisms to poultry.

Industrial field

The present invention relates to a method of administration of compositions of viable microorganisms to birds of poultry to which microorganism compositions are administered viable useful for the growth of birds, bringing the birds receive a diet with high productivity while reduce the amounts of medications, such as antibiotics, antibacterial agents and the like, or are not administered medicines.

Technical background

In feeding poultry, such as broiler chickens and the like, an intensive breeding of 40 to 60 birds per 1 tube (approximately 3.3 m2) is generally carried out in a poultry farm with the In order to reduce production costs. In addition, poultry feed for rapid growth, a daily gain in body weight of 40 to 55 g per day on average, so that the birds can be sent for sale in approximately 2 months after incubation. Under these conditions, the bird is always under enormous stress, and often sick and diseases spread among poultry. When the feed is thrown to the ground, since the birds bite the food and the like, which is mixed with the excrements of the soil, the toxic bacteria spread very easily to all the birds that are in the feeding grounds. In addition, in an open feeding installation, or even in some cases in a windowless feeding installation, toxic bacteria can spread through the feeding enclosure. In addition, for example, even if the chicken pen is disinfected each time the chickens are sold, since small animals such as rats and the like in the surrounding areas are not affected by this treatment, these animals carry bacteria such as salmonella and similar that they transmit to the poultry when chicks are again placed in the feeding facility.

In order to maintain a high productivity, while avoiding a reduction in the rate of growth and similar during the feeding period, as well as to prevent contamination of poultry with bacteria toxic under these conditions, they are usually administered at Birds various medications, such as antibiotics, agents antibacterials and the like.

However, since there is the possibility of that residual drugs remain in the meat and eggs, and also due to the danger of generating bacterial strains resistant to medications (as has been published in many cases), has gone increasing the demand for farm products from users cattle free of drugs, which are produced without using medicines.

Moreover, in natural conditions, without use intensive breeding of a large number of birds, can not Expect high productivity. However, the poultry breeding, such as chickens and the like, without use medications such as antibiotics, agents antibacterials and the like. There is now a great demand for meat of chicken and eggs raised in natural conditions, that is, the called natural poultry, due to safety and good flavor, and they are sold in the market at a high price.

As a consequence, interest has been directed towards the development of a method of feeding in the raising of Poultry, such as chickens and the like, that does not lead to contamination with toxic bacteria, at the same time as maintains high productivity, by a free feeding method of drugs (food in which no such medications are used as antibiotics, antibacterial agents and the like, during part or throughout the feeding period, so that the medications do not remain in the birds at least at the time of your shipment for sale).

When conventional intensive rearing of a large number of birds is carried out by simply avoiding the medication administration, birds easily fit infected with toxic bacteria. Therefore, it is not possible to maintain productivity at a commercially acceptable level without any Contamination with toxic bacteria.

As a means to reduce the infection of birds with toxic bacteria without relying on medications, such as antibiotics, antibacterial agents and the like, it has been proposed to administer microorganisms and the like useful for the growth of various types of poultry. It is known that bacteria belonging to the genus Bacillus , lactic acid bacteria belonging to the genus Lactobacillus , bacteria belonging to the genus Bifidobacterium and the like are useful microorganisms for feed additives or as various EC preparations (competitive exclusion). Some of these microorganisms are commercially available as preparations of viable microorganisms of feed additives for poultry. However, although the effects are recognized to some extent, these are not enough. In particular, no means have been reported to allow drug-free poultry feeding in intensive rearing of a large number of poultry and with a commercially acceptable level, to provide these viable microorganism preparations.

For example, Japanese Patent No. 2528055 and JP-B-3-79988 (the term "JP-B" as used herein means "Japanese Patent Publication Examined") describe that the gain in body weight and the ratios Conversion of feed into animals can be improved and effects such as control action of intestinal function and the like can be obtained by administering a preparation of viable microorganisms comprising Bacillus subtilis to animals. It is possible to carry out a drug-free diet by applying the medium to, for example, broiler chickens, but only in certain limited regions where these chickens are not produced on a large scale. However, in many regions where chickens are fed frequently, since they are frequently infected with toxic bacteria that spread easily through feeding facilities as described above, it is extremely difficult to carry out a commercial drug-free diet of broiler chickens using this medium. In addition, even if the drug-free feeding of broiler chickens using this medium is possible, the rate of gain of the body weight of the birds is lower than with a diet using medications. As a result, the high productivity of the feed with applied drug is not obtained.

In addition, microorganism preparations currently viable in the market have problems, such as the reduction of the viable number during the distribution stages, the poor colonization capacity after flora formation intestinal bacterial, the need for continued administration of preparations of viable microorganisms for a period of prolonged time, and the like. In particular, the need for a continuous administration of the preparations of viable microorganisms for a prolonged period of time, due to poor colonization capacity after formation of intestinal bacterial flora, is currently a serious obstacle for practical use from an economic point of view.

Description of the invention

An object of the present invention is provide a method for the administration of compositions of viable microorganisms to poultry, so that the Bird-free drug feeding can be carried out while maintaining good productivity.

To solve the aforementioned problems, the authors of the present invention have carried out a selection analysis of useful bacteria from a wide range of natural sources and have found that certain lactic acid bacteria isolated from the intestinal tract of a chicken can colonize easily the intestinal tract of newborn chickens immediately after incubation. In addition, the authors of the present invention have found that, in particular, a heterofermentation lactic acid bacterium isolated from a chicken's ileum and a homofermentation lactic acid bacterium isolated from a chicken's cecum can easily colonize the intestinal tract of the Newborn chicks and have various effects, such as inhibition of the growth of toxic bacteria, prevention of diarrhea, growth enhancement, improvement of the breeding rate, improvement of feed conversion ratios and the like. In addition, the authors of the present invention have found that lactic acid bacteria can colonize the intestinal tract of newborn chicks immediately after birth by simply spraying a suspension of viable microorganisms of these lactobacillus lactic acid bacteria only one time to newborn chicks within four days after incubation.

Furthermore, the authors of the present invention have found that, when lactic acid bacteria are used whose colonization takes place in combination with the administration of viable microorganisms of Bacillus subtilis during the period from the newborn breeding stage to the mature bird stage in Poultry-free drug feeding, body weight gain in the initial feeding stage can be improved and a sufficient breeding rate can be obtained compared to the single administration of viable microorganisms of Bacillus subtilis , so that A drug-free feeding of poultry can be carried out in intensive rearing of a large number of birds with high commercially profitable productivity. The present invention has been carried out on the basis of these findings.

According to this, the present invention relates to a method for the administration of viable microorganisms to Poultry comprising:

administer to the poultry a first composition comprising viable bacteria belonging to Lactobacillus reuteri and Lactobacillus johnsonii , at the stage of newborn pups.

In addition, the present invention relates to a method for administration of microorganism compositions viable for poultry, comprising:

administration of a composition of viable microorganisms for poultry comprising viable lactic acid bacteria belonging to Lactobacillus reuteri and Lactobacillus johnsonii at the stage of newborn pups; Y

administration of a composition of viable microorganisms for poultry comprising a viable microorganism belonging to Bacillus subtilis .

The best way to carry out the invention

The method of the present invention for administering viable microorganism compositions for poultry includes the administration of a viable microorganism composition for poultry comprising viable lactic acid bacteria belonging to Lactobacillus reuteri and Lactobacillus johnsonii (to be cited from hereinafter as a "composition of viable microorganisms of lactic acid bacteria") to newborn pups.

The lactic acid bacteria described above belonging to the genus Lactobacillus are facultative anaerobic bacteria. Not only those isolated from the intestinal tract of poultry but also other strains isolated from a natural source can be used effectively. Preferred examples of the strain belonging to Lactobacillus reuteri include Lactobacillus reuteri CP-720 (Deposit No. FERM BP-6332), Lactobacillus reuteri CP-722 (Deposit No. FERM BP-6334) and the like. A preferred example of a strain belonging to Lactobacillus johsonii includes Lactobacillus johnsoii CP-721 (Deposit No. FERM BP-6333) and the like. These three strains have been deposited at the National Institute of Bioscience and Human Technology (address 1-3 Higashi 1 chome, Taukuba-shi, Ibaraki-kan 305-8566, Japan) on April 27, 1998. Other strains of Lactobacillus are known. reuteri and Lactobacillus johnsonii , such as those described in "ATCC Bacteria and Bacteriophages", 19th edition, 1996, pages 195 and 197, incorporated herein by reference.

The bacteriological properties of Lactobacillus reuteri CP-720 and CP-722 and Lactobacillus johnsonii CP-721 are shown in Table 1.

TABLE 1

one

The composition of viable microorganisms of lactic acid bacteria may also contain heterofementation lactic acid bacteria belonging to the genus Lactobacillus other than Lactobacillus reuteri such as Lactobacillus brevis, Lactobacillus buchneri and the like, as well as homofermentation lactic acid bacteria belonging to the genus Lactobacillus. to Lactobacillus johnsonii , such as Lactobacillus gasseri, Lactobacillus crispatus and the like, belonging to the group of Lactobacillus acidophillus . Many different strains of many different genera of Lactobacillus are known, such as those described in "ATCC Bacteria and Bacteriophages" 19th edition, 1996, pages 192-199, which is incorporated herein by reference.

Among the examples of the preferred medium that can be used to grow viable bacteria microorganisms lactic acid is included a milk medium, such as milk from cow, goat milk, mare's milk and the like, skimmed milk of those and a medium for lactic acid bacteria, such as BL medium, Briggs liver broth medium, MRS medium, GAM medium, TTY medium and the like.

Lactic acid bacteria can be cultivate at 25 to 45 ° C, more preferably 30 to 40 ° C, and for 6 to 30 hours, more preferably 10 to 24 hours. The broth as well obtained can be used directly as a composition of viable microorganisms of lactic acid bacteria, storing it as such at about 5 ° C until use. Alternatively, the microorganisms can be recovered by centrifugation, mixed with  a protective agent and then lyophilized under vacuum. The dust of resulting bacteria can be stored in a cold and dark chamber and used as a composition of viable microorganisms of bacteria of lactic acid by suspension, mixing or dissolution of the powder when it's used. The powder of dried microorganisms prepared In this way it is more preferred because it resists long periods of storage.

The composition of viable microorganisms of Lactic acid bacteria may contain a carrier and diluent. The vehicle and diluent are not limited in particular, and are select between pharmaceutical vehicles and diluents or nutritionally acceptable. In addition, the composition of viable microorganisms of lactic acid bacteria can contain a feed for poultry (ration).

In the present invention, microorganisms belonging to Lactobacillus reuteri and Lactobacillus johnsonii can undergo an appropriate mutation treatment, such as exposure to ultraviolet light, X-rays or radiation, and to a chemical treatment with a mutagenic compound (eg nitrous guanidine, dye of acridine). Mutants can also be prepared by insertion, deletion or replacement of nucleotides, as well as by spontaneous mutation. The terms Lactobacillus reuteri and Lactobacillus johnsonii include these mutants.

The term "newly hatched stadium born "means the period immediately following the incubation of the offspring (for example chick, pavipollo, pigeon or similar), specifically a period of 0 to about 4 days after incubation The administration time of the composition of viable microorganisms of lactic acid bacteria is not particularly limited as long as it is within the stage of newborn pups; however, it is preferred to administer the composition within 4 days after incubation, more preferably within 2 days after incubation, a period before colonization of intestinal bacterial flora in the intestinal tract of the newborn. The number of times of administration is not particularly limited; but nevertheless, you can usually have enough effects for just one administration. The effects obtained become more stable and insurance as the number of administration times increases. However, three or more times of administration does not result economic, and the effects thus obtained are almost the same as with twice administration.

Although not limited in particular, the administration of the composition of viable microorganisms of lactic acid bacteria can be carried out by oral administration Specifically, the composition can be orally administered by its addition to drinking water or the like and let the offspring ingest it freely, or it can be sprayed from a position above the young using a sprayer or similar. The method of spray administration is the preferred due to the habit of the young to twitter while they open their peaks up, so that the composition of viable microorganisms that has a high concentration can orally administered at the stage of newborn pups so easy and safe

In carrying out the administration, the density total lactic acid bacteria in the composition of viable microorganisms of lactic acid bacteria is preferably from 10 6 to 10 10 viable microorganisms per gram, more preferably 10 7 to 10 9 microorganisms viable per gram. In addition, the composition of microorganisms viable lactic acid bacteria is preferably administered in an amount of 1x10 3 to 1x10 10 microorganisms viable for breeding, more preferably 1x10 4 to 1x10 6 of viable microorganisms by breeding.

In the method of the present invention, the composition of viable microorganisms for poultry comprises a viable microorganism belonging to Bacillus subtilis (hereafter referred to as "composition of viable microorganisms of Bacillus subtilis ") is administered in addition to the administration of the composition of viable microorganisms of lactic acid bacteria described above.

Preferred examples of the Bacillus subtilis strain include Bacillus subtilis C-3102 (Deposit No. FERM BP-1096) that has been deposited with the Institute of Bioscience and Human Technology (former name: Fermentation Research Institute), Agency of Industrial Science and Technology (Address: 1-3, Higashi 1 chome, Tsukuba-shi (former address: Yatabe-machi, Tsukuba gun), Ibaraki-ken 305-8566 (old code in force: 305), Japan) on June 28, 1986, and the like. The bacteriological properties of Bacillus subtilis C-3102 are already described in Japanese Patent No. 2528055, JP-B-3-79988 and US Patent Re. 34,837. Many different strains of Bacillus subtilis are known , such as those described in "ATCC Bacteria and Bacteriophages", 19th edition, 1996, pages 57-63, which is incorporated herein by reference.

When growing Bacillus subtilis C-3102, an aqueous or solid medium containing materials, such as carbon sources, nitrogen sources, inorganic substances and the like, which are generally used in the cultivation of microorganisms, can be used as a culture medium As a culture medium. Examples of carbon sources include those that can be assimilated, such as glucose, fructose, sucrose, starch, molasses and the like. Examples of nitrogen sources include peptone, meat extract, casein hydrolyzate, ammonium sulfate and the like. As the case demands, you can also add, as inorganic components, salts of magnesium, potassium, sodium, calcium, iron, manganese and the like, vitamins, amino acids, antifoam agents, surfactants and the like. Cultivation is preferably carried out aerobically. The starting pH of the medium is preferably 5 to 9, more preferably 6 to 8; the culture temperature is preferably 20 to 50 ° C, more preferably 35 to 40 ° C; and the cultivation period is preferably 12 hours to 7 days.

The culture mixture obtained in this way can be used as the viable microorganism composition of Bacillus subtilis as such or as its concentrated product or as isolated cells thereof, directly or after adding additives such as fillers and the like. Loads are not limited in particular, and examples include calcium carbonate, degreased rice bran, corn grits, cornmeal, wheat bran, skimmed milk powder and the like.

The viable microorganism composition of Bacillus subtilis may contain a carrier or diluent. The carrier and diluent are not limited in particular, and are selected from pharmaceutically or nutritionally acceptable carriers and diluents. In addition, the composition of viable microorganisms of Bacillus subtilis may contain a poultry feed.

In the present invention, the microorganism belonging to Bacillus subtilis can be subjected to an appropriate mutation treatment, such as exposure to ultraviolet light, X-rays or radiation, and a chemical treatment with a mutagenic compound (eg, nitrosoguanidine, acridine dye ). Mutants can also be prepared by insertion, deletion or replacement of nucleotides, as well as spontaneous mutation. The term Bacillus subtilis includes these mutants.

The administration time of the composition of viable microorganisms of Bacillus subtilis is not limited in particular; however, to obtain an appropriate colonization of intestinal bacterial flora, it is preferable to administer the composition of viable microorganisms of Bacillus subtilis after administration of the composition of viable microorganisms of lactic acid bacteria described above. Also, in the case of a feeding method in which antibiotics, antibacterial agents and the like are not administered for a period of time between the stage of newborn pups and the termination stage, it is preferable to administer the composition of viable microorganisms of Bacillus subtilis during this time. In the case of feeding broiler chickens, for example, it can be administered during an optional period between the stage of newborn pups and the termination stage; however, when a drug-free diet is carried out only during the stage of greatest growth and the termination stage so that the drugs do not remain in the body of the broiler chicken at the time of shipment of the chickens for sale, the Composition of viable microorganisms of Bacillus subtilis can be administered during the growth stage and the termination stage.

The administration of the composition of viable microorganisms of Bacillus subtilis can be carried out by oral ingestion after its addition to the feed, drinking water or the like.

For example, when the composition of viable microorganisms of Bacillus subtilis containing Bacillus subtilis C-3102 is mixed with feed at the time of administration, it is preferred that the feed has a density of microorganisms from 10 5 to 10 8} viable microorganisms per gram in the form of spores and / or plant cells.

The method of the present invention for administration of the compositions of viable microorganisms for Poultry can be applied not only to broiler chickens to which it is fed in intensive breeding regime with a large number of birds but also to broiler chickens that are fed in other conditions and other domestic birds such as ducks, geese, quail, wild ducks, ostriches and the like, as well as to companion birds and the like.

When the microorganism compositions viable for poultry are administered by the method before described, an intestinal bacterial flora can be formed in which the bacteria useful for the bird growth, so that birds grow healthy. Be can obtain various effects, such as growth inhibition of toxic bacteria, diarrhea prevention, potentiation of growth, improvement of feed conversion ratios and similar, and drug-free feeding of The chicken broilers. In addition, since useful bacteria dominate the intestinal bacterial flora, the poultry grows with good health, so you can get delicious eggs, meat and similar in the case of edible birds due to the quality remarkably improved products. In addition, meat, eggs and Similar are safe and are not contaminated with toxic bacteria They poison food.

Examples

Having described in general terms this invention, can be understood even better with reference to certain specific examples given here for purposes illustrative only and not to be construed as limiting unless otherwise specified.

Example 1 of Production

A portion of 4.5 g of skim milk was dissolved in 50 g of water, pasteurized at 100 ° C for 10 minutes and then cooled to room temperature. A loop of Lactobacillus reuteri CP-720 was inoculated into the solution thus prepared and statically cultured for 24 hours at 37 ° C to obtain a first initiator (lactic acid bacteria: 1x10 9 viable microorganisms per gram). Then 15 g of the first initiator were inoculated into 500 g of skim milk (solids content, 9% by weight) that had been pasteurized at 90 ° C and cultured for 20 hours at 37 ° C to obtain a second initiator. The second initiator contained 2x108 viable microorganisms per gram.

It was placed in a bucket fermenter, a medium prepared by dissolving 200 g of casein peptone, 200 g of yeast extract, 100 g of sodium citrate, and 200 g of glucose in 20 kg of water and adjust the pH of the medium to 7.0 with solution of 1 N sodium hydroxide, was pasteurized at 95 ° C for 15 minutes and was then cooled to room temperature. They were inoculated after 3 parts by weight of the second initiator described above in 100 parts by weight of the medium and cultured statically for 20 hours at 37 ° C

The culture broth thus obtained was centrifuged to recover the microorganisms that were subsequently lyophilized using, as dispersion medium, 1 kg of solution containing 10% by weight of skim milk and 1% by weight of sodium glutamate, which had been pasteurized. at 90 ° C previously, thus obtaining 146 g of powder from viable microorganisms of Lactobacillus reuteri CP-720. The viable microorganism powder contained 7x10 10 viable microorganisms per gram.

Example 2 of Production

One medium, prepared by dissolving 200 g of ox peptone, 60 g of soy peptone, 100 g of yeast extract, 100 g of sodium acetate, 40 g of dipotassium phosphate, 60 g of diamonium citrate and 400 g of glucose in 20 kg of water and adjusting the pH of the medium to 7.0 with 1 N sodium hydroxide solution, placed in a cuvette fermenter, pasteurized at 95 ° C for 15 minutes and then cooled to room temperature. 500 g of a second Lactobacillus johnsonii CP-721 initiator which had previously been pre-cultured by the same procedure as Production Example 1 in the medium was then inoculated to perform a stationary culture of 18 hours at 35 ° C.

The culture broth thus obtained was centrifuged to collect the microorganisms that were subsequently lyophilized using as a dispersion medium a solution containing 10% by weight of skim milk and 1% by weight of sodium glutamate, which had been pasteurized at 90 ° C previously, thereby obtaining 141 g of powder from viable microorganisms of Lactobacillus johnsonii CP-721. The microorganism powder contained 6.5x1010 viable microorganisms per gram.

Example 3 of Production

A second Lactobacillus reuteri CP-722 initiator prepared by the same procedure of Production Example 1 was inoculated in a medium prepared and pasteurized in the same manner as described in Production Example 1 which had the same composition and component weights , and was statically cultured for 20 hours at 37 ° C. The culture broth thus obtained was centrifuged to recover the cells that were subsequently lyophilized, using as a dispersion medium, a solution containing 10% by weight of skim milk and 1% by weight of sodium glutamate, which had been pasteurized at 95 ° C. previously, thereby obtaining 151 g of powder from viable microorganisms of Lactobacillus reuteri CP-722. The viable microorganism powder contained 6.8x10 10 viable microorganisms per gram.

Example 4 of Production

Viable microorganism powders obtained in Production Examples 1 to 3 were combined in 1 part portions and carefully mixed with 7 parts of dextrin to obtain a viable microorganism composition of lactic acid bacteria for poultry containing three strains of Lactobacillus . The composition of viable microorganisms contained 2,0x10 10 viable microorganisms per gram.

Production Example 5

One medium, prepared by dissolving 200 g of soy peptone, 10 g of dipotassium phosphate and 200 g of molasses in 10 kg of water and adjusting the pH of the medium to 7.5 with 1 N sodium hydroxide solution, placed in a cuvette fermenter, pasteurized at 95 ° C for 60 minutes and then cooled to 37 ° C. 100 g of Bacillus subtilis C-3102 culture broth, which had been precultured in advance, was then inoculated in the medium and stirred aerobically for 40 hours at 37 ° C. The culture broth thus obtained was centrifuged to recover the microorganism that was subsequently mixed with the same weight of skim milk and lyophilized in vacuo, thereby obtaining 750 g of the composition of viable microorganisms of Bacillus subtilis for poultry. The composition of viable microorganisms of Bacillus subtilis for poultry contained 1,2x10 10 viable microorganisms per gram.

Example 1 Field I trial of the effect (drug-free trial at the stage of higher growth and later stage)

The test was carried out using Chunky (Brand name, commercial line). Chicks were given by spraying (0 days after incubation), placed in a chicken cage, 12 g of the composition of viable microorganisms of lactic acid bacteria for poultry prepared in Production Example 4 (number 4). Total lactic acid bacteria: 2.0x10 10 viable microorganisms per gram) that had been uniformly suspended in 4 kg of pure water. As feed for provision, "Pre-initiation feed for chickens (Hina Ezuke)" was used for use as starter feed (containing antibiotic) and "Initiation AT (Zenki AT)" for use at the initiation stage (containing antibiotic) at the beginning feeding stage (0 to 5 days after incubation) and initiation stage (5 to 21 days after incubation), respectively. "Brogoal A" (not containing antibiotics) was used for use in the termination stage and which had been mixed with 9x105 cells / gram of the composition of viable microorganisms of Bacillus subtilis for poultry prepared in the Example 5 of Production, during the period of the fastest growing stage (from 21 days to 43 days after incubation) to the termination stage (from 43 days a day of shipment for sale) (all these commercial feed items were manufactured by Chubu Shiryo). Other feed management was carried out in accordance with the method of feeding the chickens, which is conventionally carried out in poultry farms. The test results are shown in Table 2.

Example 1 Comparative

Feeding was carried out in the same manner as in Example 1, except that administration of the composition of viable microorganisms of lactic bacteria was not performed, a "Product of the highest growth AT (Kohki AT)" was used for use in the fastest growing stage (which contained antibiotics) (manufactured by Chubu Shiryo, Co.) as I think of the highest growing stage instead of "Brogoal A" for termination use, and the composition of viable Bacillus microorganisms was not provided subtilis throughout the entire stages. The results are shown in Table 2. In this regard, the "product of the highest growth AT (Kohki AT)" is an article of a feed that has the same composition as "Brogoal A" except in antibiotics.

TABLE 2

2

         \ dotable {\ tabskip \ tabcolsep # \ hfil \ + # \ hfil \ tabskip0ptplus1fil \ dddarstrut \ cr} {
 * \ + Breeding rate (the number of birds sent for sale and the
number of birds of the game) x 100 \ cr ** \ +
 \ begin {minipage} [t] {145mm} Feed conversion ratio:
total amount of feed ingested during feeding (g) / total
of body weight gain (g) x 100 \ end {minipage} \ cr ***
\ + \ begin {minipage} [t] {145mm} Production score: (weight
average body x parenting rate / conversion ratio of
food x age in days when sent for sale) x
100 \ end {minipage} \ cr}
      

The value of the average body weight gain in Example 1 it was lower than that in Comparative Example 1 in a factor of approximately 1%, which was assumed due to the quality of The chicks tested. That is, the body weight of Chicks were normal (41.9 g / chicken) in Comparative Example 1 but it was closer to that of the young chicks (38.4 g / chicken) of Example 1, so that its growth became inferior. Given the other results of Example 1 were superior to those of Example Comparative 1, productivity was improved.

Example 2 Field II trial of the effect (drug-free trial at the stage of initiation and after the initiation stage)

The test was carried out using Chunky (brand name, commercial line). The chicks (on day 0 after incubation), placed in a chicken cage, received, by spraying, 12 g of the composition of viable microorganisms of lactic acid bacteria for birds prepared in Production Example 4 (total number of lactic acid bacteria; 2.0 x 10 10 viable microorganisms per gram) that had been uniformly suspended in 4 kg of pure water. As I think of provision, the "Chick Prestarter Feed" was used for use as starter feed at the start feeding stage (from 0 to 5 days after incubation). The "initiation AT (Zenki AT)" for use in the initiation stage from which the antibiotics had been removed and the "Brogoal A" for termination use, which had been mixed with 9x105 viable microorganisms per gram of the Composition of viable microorganisms of Bacillus subtilis for birds prepared in Production Example 5, were used at the initiation stage (5 to 21 days after incubation) and during the period of the highest growth stage (from 21 days to 43 days after incubation) to the termination stage (from 43 days to the shipping stage for sale), respectively. Other feeding management was carried out following the chicken feeding method conventionally carried out in poultry farms. The test results are shown in Table 3.

Example 2 Comparative

Feeding was carried out in the same manner as described in Example 2, except that the "Initiation AT (Zenki AT)" for use in the initial stage and the "Highest growing AT (Kohki AT)" to The highest growth stage was used in the initial stage and the highest growth stage respectively, and the composition of viable microorganisms of Bacillus subtilis was not provided throughout the entire stages. The results are shown in Table 3.

         \ vskip1.000000 \ baselineskip
      

TABLE 3

3

Since drug-free feed was used in initiation stage and after initiation in Example 2, while in Comparative Example 2 the feed was used that It contained drug, body weight gain in the first case It was slightly lower than the value of 51.6 g / day. Chicken, but the rate of upbringing was 97%, which is an excellent fact and the production score was 244, showing results of An economic production.

Example 3 Field III trial of the effect (drug-free trial)

The test was carried out using Chunky (brand name, commercial line). Chicks were given by spraying (day 0 after incubation), placed in a chicken cage, 40 g of the composition of viable microorganisms of lactic acid bacteria for poultry prepared in Production Example 4 (number Total lactic acid bacteria: 2x10 10 viable microorganisms per gram) that had been uniformly suspended in 4 kg of pure water. The "AT of initiation (Zenki AT)", from which the antibiotics had been removed, for the initiation stage and the "Brogoal A" for termination use, which had been mixed with 9x10 5, were used as the provision feed. Viable microorganisms per gram of the composition of viable microorganisms of Bacillus subtilis for poultry prepared in Production Example 5, during the period from the feeding stage of the beginning to the initiation stage (from 0 to 21 days after incubation) and during the period of the stage of greatest growth to the stage of completion (from 21 days a day of shipment for sale), respectively. Another feeding management was carried out in accordance with the conventional chicken feeding method in poultry farms. The test results are shown in Table 4.

Example 3 Comparative

Feeding was carried out in the same manner as described in Example 3, except that "Initiation AT (Zenki AT)" was used for use in the initiation state, "The fastest growing AT (Kohki AT) "for the fastest growing stage and the" Brogoal A "for termination use, respectively, as I think during the period from the feeding stage from the beginning to the initiation stage, in the fastest growing stage (from 21 to 43 days after incubation) and at the termination stage (43 days per day of shipment for sale), respectively, and the composition of viable microorganisms of Bacillus subtilis was not provided throughout the entire stages. The results are shown in Table 4.

TABLE 4

4

The test of Example 3 was not carried out at same time as the test of Comparative Example 3 but after if the feeding of Comparative Example 3 is completed, and, given that there was a cold blow during the feeding period of the Example 3, the final growth and the breeding rate in Example 3 Comparative were influenced by the cold wave. But nevertheless, although the feeding was carried out without administration of antibiotics and antibacterial agents throughout the period complete, the production results of Example 3 were identical to those of Comparative Example 3.

In addition, the comparative tests corresponding to Examples 1 to 3 were not performed with a feed carried out without drugs and without administering the composition of viable microorganisms of lactic acid bacteria for birds and the composition of viable microorganisms of Bacillus subtilis for birds of corral, because you can't get normal growth under these conditions.

Example 4

In Example 3 and Comparative Example 3, recent feces were collected from chickens the day before being sent for sale and, the excrements of three birds were used as a sample, the group of coliform bacteria and bacteria of the genus Salmonella were measured in the faeces. The results are shown in Table 5. In this case, the serotype of each bacterium of the genus Salmonella was 07.

TABLE 5

5

In comparison with Comparative Example 3, the group of coliforms decreased significantly in Example 3, and the detection ratio of bacteria of the genus Salmonella was also significantly reduced.

Example 5

A comparison was made about the sensation at palate of the chicken meat raised in Example 3 and Example 3 Comparative. Since a trial cannot be carried out precise sensory when the meat itself is tested Due to several treatment factors, a soup of Complete chicken and underwent sensory testing. It was used in the rehearse the edible part that remains after sacrifice, bleeding  and subsequent removal of feathers, heads, legs and organs (gutting III), as a whole chicken and mixed a part by weight of the sample with 4 parts by weight of water and 1.8% salt and boiled for 2 hours After filtering the mixture through cotton bleach, the filtrate (soup) was prepared and the Concentration of the soup to three times the weight of the chicken. The soup samples thus obtained were subjected to sensory testing in the one that 38 panelists could select the flavor sample more delicious. The results are shown in Table 6.

TABLE 6

6

In comparison to the chicken in Example 3 Comparative, the chicken of Example 3 was selected by the sensation that produced on the palate by a large number of panelists, thus confirming the delicious nature of the chicken produced with a drug free food.

In a taste evaluation carried out at same time in connection with this, good evaluations were obtained  for chicken soup from Example 3, statistically significant with a level of significance of 1%, on each of the points: (1) preference, (2) richness of taste and (3) taste suitable.

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Example 1 of reference

Completely drug-free assay, only composition of viable microorganisms of Bacillus subtilis for poultry

The test was carried out using Chunky birds from a district where the broiler chickens were fed in large numbers. As I think of provision, "Gold initiation crumbs for broiler chickens (Broiler Gold Zenki Crumble)" were used from which antibiotics had been removed and "Product for last stage for chickens (Broiler Shiage)" (which did not contain antibiotics or similar) (both manufactured by Shikoku Haigo Shiryo, Co.), each of which had been mixed with 1x10 6 viable microorganisms per gram of composition of viable microorganisms of Bacillus subtilis for poultry prepared in Production Example 5 and enriched with vitamins and minerals, during the period from the feeding stage from beginning to the initiation stage (from 0 to 21 days after incubation) and during the period from the stage of greatest growth to the finishing stage (from 21 days after incubation on the day of shipment for sale), respectively. Other feed handling was carried out in accordance with the conventional method of chicken feeding carried out in poultry farms. The test results are shown in Table 7.

Example 4 Comparative

Feeding was carried out in the same manner as in Reference Example 1, except that "Gold Starter Crumbs for Chickens""(Broiler Gold Zenki Crumble)" (containing antibiotics), "Broiler S" containing antibiotics and "Broiler Finisher (Broiler Shiage)" each manufactured by Shikoku Haigo Shiryo, Co.) as I think during the period of the feeding stage from the beginning to the initiation stage, in the stage of greatest growth (of 21 32 days after incubation on the day of shipment for sale), and at the termination stage (32 days after incubation on the day of shipment for sale), respectively, and the composition of viable Bacillus microorganisms was not provided subtilis The results are shown in Table 7.

TABLE 7

7

A higher breeding rate was obtained in the Reference Example 1 than in Comparative Example 4, but the gain in body weight at the stage of initiation of power was delayed and the delay left a mark until the final, so that the weight gain was low through of the feeding period as a result and it was not reached a commercially acceptable level.

Nor was the drug-free breeding trial provided by providing only the composition of viable microorganisms of lactic acid bacteria for poultry without administration of the composition of viable microorganisms of Bacillus subtilis for poultry, because this test can only be carried out. carried out in a place where there is no environmental contamination such as in an experimental installation under complete sanitary conditions but not in the field of intensively exploited poultry farms.

Industrial applicability

Since the administration of a viable microorganism composition for poultry comprising viable specific lactic acid bacteria is carried out at a specific stage in combination with the administration of a viable microorganism composition for poultry comprising viable Bacillus subtilis , the method of the present invention for the administration of compositions of viable microorganisms for birds can exert various effects, such as inhibition of the growth of toxic bacteria, prevention of diarrhea, growth enhancement, improvement of feed conversion ratios and the like. , also making possible drug-free feeding for chickens with high productivity and, particularly in the case of edible chickens, producing delicious and safe eggs and meat with high productivity. Accordingly, the method of the present invention for the administration of compositions of viable microorganisms for birds is useful for feeding edible birds, such as chickens, particularly broiler chickens, and the like.

Claims (16)

1. Use of a composition of viable microorganisms for the preparation of a pharmaceutical composition for inhibition of the growth of toxic bacteria, prevention of diarrhea and / or enhancement of poultry growth, where the first composition comprising bacteria is administered to the birds viable belonging to Lactobacillus reuteri and Lactobacillus johnsonii in the stage of newborn pups and where the said birds are administered a second composition comprising a viable microorganism belonging to Bacillus subtilis . 2. A use according to claim 1, where said first composition is administered within four days after the birds have been incubated 3. The use according to claim 1, where said first composition is administered within 2 days after incubation of said birds. 4. The use according to any of claims 1 to 3, wherein said Bacillus reuteri is at least one member selected from the group consisting of

(to)

Lactobacillus reuteri CP-720, Deposit No. FERM BP-6332, and a mutant thereof,

(b)

Lactobacillus reuteri CP-722, Deposit No. FERM BP-6334, and a mutant thereof.

5. The use according to any of claims 1 to 4, wherein said Lactobacillus johnsonii is Lactobacillus johnsonii CP-721, Deposit No. FERM BP-6333, or a mutant thereof. 6. The use according to any of the claims 1 to 5, wherein said first composition and / or the said second composition further comprises a vehicle or diluent 7. The use according to any of the claims 1 to 6, wherein said first composition and / or the said second composition further comprises a feed for birds of corral. 8. The use according to any of the claims 1 to 7, wherein said first composition and / or the Said second composition is administered orally. 9. The use according to any of the claims 1 to 7, wherein said first composition is administered by spraying. 10. The use according to any of the claims 1 to 9, wherein said first composition it comprises 10 6 to 10 10 of said bacteria per gram. 11. The use according to any of the claims 1 to 10, where they are administered to poultry 1x10 3 to 1x10 10 of those cited. 12. The use according to any of the claims 1 to 11, wherein said first composition is Administer only once or twice. 13. The use according to any of claims 1 to 12 wherein said Bacillus subtilis is Bacillus subtilis C-3102, Deposit No. FERM BP-1096, or a mutant thereof. 14. The use according to any one of claims 1 to 13, wherein said second composition comprises from 10 5 to 10 8 of said viable Bacillus subtilis per gram. 15. The use according to any of the claims 1 to 14, wherein said second composition is administers after the administration of said first composition. 16. The use according to any of the claims 1 to 15, wherein said bird is a chicken, duck, Goose, quail, wild duck, ostrich or companion bird.